Inhomogeneity Based Characterization of Distribution Patterns on the Plasma Membrane
Fig 3
Detecting protein distributions using widefield microscopy.
(A) Supported plasma membrane sheet of a MEF cell with Na+/K+ ATPase antibody staining. (B) Zoomed in region highlighted by the yellow rectangle in (A). (C) Plasma membrane sheet of a MEF cell exhibiting TfR antibody staining. (D) Zoomed in region highlighted by the yellow rectangle in (C). (E) Green: DiO staining for the MEF cell isolated plasma membrane. Red: Caveolin-1 antibody staining. (F) Zoomed in region highlighted by the yellow rectangle in (E). (G) The comparison with the analysis of simulated images suggests Na+/K+ ATPase was randomly distributed, TfR was organized into clusters, and Cav1 showed a polar distribution of clusters, as expected. (H) The inhomogeneity measure reveals a homogeneous distribution of Na+/K+ ATPase (deviation from random: -0.02) and increasing inhomogeneous organization for TfR (deviation from random: 0.51) and Cav1 (deviation from random: 1.1). Scale bars in (A, C, and E): 5 μm. Scale bars in (B, D, and F): 3 μm. Error bars represent the SEM. Number of analyzed images for Na+/K+ ATPase is 41, TfR is 47, and Cav1 is 50. The average r2 is 0.92, 0.93, and 0.93, respectively.